Source: KANSAS STATE UNIV submitted to NRP
BACULOVIRUS GENE REGULATION AND REPLICATION
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1000348
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Oct 1, 2013
Project End Date
Sep 30, 2018
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
KANSAS STATE UNIV
(N/A)
MANHATTAN,KS 66506
Performing Department
Biology
Non Technical Summary
We are studying the function of two insect viral genes. One gene, ac79, may be involved in DNA repair. The other gene, ac92, may be involved in stabilizing the viral 3D structure. These genes may be manipulated to virulence. Insect viruses are non-toxic alternatives to chemical pesticides and can be improved as such.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2153110106050%
2154099106050%
Knowledge Area
215 - Biological Control of Pests Affecting Plants;

Subject Of Investigation
4099 - Microorganisms, general/other; 3110 - Insects;

Field Of Science
1060 - Biology (whole systems);
Goals / Objectives
Define the functional mechanisms and intermediary factors of two conserved insect virus genes during virus replication.
Project Methods
In general, we will use classical molecular biology, virology and biochemical methods. These include the constructions of recombinant viruses with site directed mutations in putative crucial motifs of the genes of interest. The phenotype of recombinant viruses will be evaluated in cell culture by assessing viral replication in different cell lines. The function of the wild-type or altered gene product of interest will be analyzed in vitro. Host or viral targets of the factors being studied will be screened. The identification of the targets will provide clues to gene function.

Progress 10/01/13 to 09/30/18

Outputs
Target Audience:Target audiences include several levels of research scientists-graduate students, undergraduate students and a research assistant. All have been part of the project. Results from the project have been discussed at the annual Microbe meeting of the American Society for Microbiology, at the annual meeting of the Society for Invertebrate Pathology and at local and regional state-wide meetings. Thus, those attending these meetings are also target audiences. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Baculoviruses are insect viruses that can infect and kill insects that eat and destroy agricultural products and forrests. Since baculoviruses have been used as biological pest control agents, learnign about the genes that allow their replication or interact with the insect host is important to imporve their efficacy. We further characterized a baculovirus sulfhydryl oxidase gene, developed a method for our system to identify the substrates and identified some substrates of the enzyme. The role of these substrate genes is being further studied. Some arboviruses are transmitted via mosquitoes and can infect livestock when an infected mosquito takes a blood meal. Aedes aegypti is being used in our studies. This may affect agriculture and further transmission of diseases. This mosquito species is the vector of chikungunya, dengue and Zika. We are studying the sites of replication of the arbovirus Sindbis virus in Ae. aegypti and requirements for replication at the primary site of infection. In addition, we are studying the role of the mosquito fibroblast growth factor gene in helping the virus escape the primary site of replication.

Publications


    Progress 10/01/16 to 09/30/17

    Outputs
    Target Audience:Target audiences include graduate and undergraduate students, postdoctoral fellow scientists, and research assistants, who have been involved in the project. Results from the project have been discussed at the annual meetings of the American Society for Virology and the Society for Insect Pathology, so members of these societies are also target audiences. The readership of virology journals where the work has been published is also a target audience. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

    Impacts
    What was accomplished under these goals? Baculoviruses are insect viruses that often target insects that consume and destroy agricultural products. Thus, baculoviruses are natural insecticides. Information regarding their replication is useful to improve these biological insecticides. We characterized a pathway necessary for baculoviruses to establish systemic infections following oral infection, the normal route of infection. To broaden our knowledge about enzymes involved in crossing the midgut of an insect via the basal lamina, we characterized matrix metalloproteases in the yellow fever mosquito, Aedes aegypti. Ae. aegypti is the vector of viruses including chikgungunya, dengue and Zika. We characterized nine matrix metalloproteases, activity and expression profiles during mosquito development and chikungunya infection. We looked at the role of baculovirus genes in determining host range. Learning about host range expansion can help target more insect pests. We also characterized a baculovirus sulfhydryl oxidase gene and developed a method for our system to identify the substrates and how they affect virus assembly.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2017 Citation: Kantor, A. M., S. Dong, N. L. Held, E. Ishimwe, A. L. Passarelli, R. J. Clem, A. W. E. Franz. 2017. Identification and initial characterization of matrix metalloproteinases in the yellow fever mosquito, Aedes aegypti. Insect Mol. Biol., 26:113-126. DOI: 10.1111/imb.12275.
    • Type: Journal Articles Status: Published Year Published: 2016 Citation: Wu, C., Deng, Z., Long, Z., Cai, Y., Ying, Z., Yin, H., Yuan, M., Clem, R.J., Yang, K. and Y. Pang. 2016. Generating a host range-expanded recombinant baculovirus. Scientific Reports. 6:28072. doi: 10.1038/srep28072.
    • Type: Journal Articles Status: Published Year Published: 2016 Citation: Ardisson-Araujo, D., Lima, R., Melo, F., Clem, R.J., Huang, N., Bao, S., Sosa-Gomez, D. and B.M. Ribeiro. 2016. Genome sequence of Perigonia lusca single nucleopolyhedrovirus: insights into the evolution of a nucleotide metabolism enzyme in the family Baculoviridae. Scientific Reports. 6:24612. doi: 10.1038/srep24612.


    Progress 10/01/15 to 09/30/16

    Outputs
    Target Audience:The target audience includeds students and trainees that have worked in the projects, undergraduate students, graduate students, postdoctoral fellow, research assistant. Outcomes from the project have been disseminated at international, national, and local meetings, extending the target audience to members of the societies of these annual meeings (scientists at all levels and students). The work has been published in virology and other journals, reaching the readership of these journals. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

    Impacts
    What was accomplished under these goals? We have been working with a baculovirus gene that is a functional sulfhydryl oxidase. These genes catalyze the formation of disulfide bonds in proteins. Sulfhydryl oxidases are not common in viruses and their function during virus replication is not known. We hypothesize that the enzyme stablilizes the viral particle. Lack of activity results in non-infectious viruses and defects in assembly. Since baculoviruses are important to control agriculatural insect pests, studying the requirements for virus stability will be important to develope more environmentally stable pest control agents. We have developed a method to identify the substrates of the viral sulfhydryl oxidase. These substrates have conserved cysteines (targetted amino acids) and are conserved genes. This suggests that formation of disulfide bonds is important for viron infectivity.

    Publications


      Progress 10/01/14 to 09/30/15

      Outputs
      Target Audience:Target audiences include graduate and undergraduate students, postdoctoral fellow scientists, and research assistants, who have been involved in the project. Results from the project have been discussed at the annual meetings of the American Society for Virology and the Society for Insect Pathology, so members of these societies are also target audiences. The readership of virology journals where the work has been published is also a target audience. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

      Impacts
      What was accomplished under these goals? Baculoviruses are insect viruses that often target insects that consume and destroy agricultural products. Thus, baculoviruses are natural insecticides. Information regarding their replication is useful to improve these biological insecticides. We characterized a pathway necessary for baculoviruses to establish systemic infections following oral infection, the normal route of infection. The requirement for genes and physiological changes involved in this pathway (i.e., viral fibroblast growth factor and tracheal basal lamina remodeling) were evaluated following infection of insects by virus injection, bypassing host barriers encountered by the virus after ingestion. We found that tracheal basal lamina remodeling was not affected if the virus was delivered directly into the hemocoel. The pathology and infection processes of baculoviruses are important to develop improved pesticides. We also characterized a baculovirus sulfhydryl oxidase gene and found that it is not determining a virion phenotype in two groups of baculoviruses, single versus multiple nucleopolyhedroviruses. This study ruled out the sulfhydyl oxidase gene as the determinant of this virion phenotype. We also characterized a granulovirus matrix metalloprotease and found that it had a role in augmenting virulence and functions, at least in part, like the viral cathepsin, an enzyme necessary for virus release (virus transmission) from the dead insect.

      Publications

      • Type: Journal Articles Status: Published Year Published: 2014 Citation: Passarelli, A. L. and J. C. Means. 2014. Intrahaemocoelic infection of Trichoplusia ni with the baculovirus Autographa californica M nucleopolyhedrovirus does not induce tracheal cell basal lamina remodeling. J. Gen. Virol. 95:719-723.
      • Type: Journal Articles Status: Published Year Published: 2014 Citation: Clem, S. A., W. Wu, and A. L. Passarelli. 2014. The Trichoplusia ni single nucleopolyhedrovirus tn79 gene encodes a functional sulfhydryl oxidase enzyme that is able to support the replication of Autographa californica multiple nucleopolyhedrovirus lacking the sulfhydryl oxidase ac92 gene. Virology 460-461:207-216.
      • Type: Journal Articles Status: Published Year Published: 2015 Citation: Ishimwe, E., J. J. Hodgson, A. L. Passarelli. 2015. Expression of the Cydia pomonella granulovirus matrix metalloprotease enhances Autographa californica multiple nucleopolyhedrovirus virulence and can partially substitute for viral cathepsin. Virology 481:166-178.
      • Type: Journal Articles Status: Published Year Published: 2015 Citation: Ishimwe, E., J. J. Hodgson, R. J. Clem, and A. L. Passarelli. Reaching the melting point: 2015. Degradative enzymes and protease inhibitors involved in baculovirus infection and dissemination. Virology 479-480:637-649.


      Progress 10/01/13 to 09/30/14

      Outputs
      Target Audience: The target audiences of this project are undergraduate and graduate students, and postdoctoral fellow scientists. Results from the project have been preented at the annual meetings of the American Society for Virology and the Society for Invertebrate Pathology, making members of these societies target audience. The readership of virology journals where the work has been published is also considered target audience. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

      Impacts
      What was accomplished under these goals? Baculoviruses are insect viruses that often target insects that consume crops. Baculoviruses are natural insecticides. Information regarding their replication is useful to improve these biological pesticides. We defined the role of baculovirus genes during pathogenesis in the insect Trichoplusia ni. In addition, we characterized the function of another baculvirus gene with homology to sulfhydryl oxidases and its ability to affect viral morphogenesis.

      Publications

      • Type: Journal Articles Status: Published Year Published: 2014 Citation: Passarelli, A. L. and J. C. Means. 2014. Intrahaemocoelic infection of Trichoplusia ni with the baculovirus Autographa californica M nucleopolyhedrovirus does not induce tracheal cell basal lamina remodeling. J. Gen. Virol. 95:719-723.
      • Type: Journal Articles Status: Published Year Published: 2014 Citation: Clem, S. A., W. Wu, and A. L. Passarelli. 2014. The Trichoplusia ni single nucleopolyhedrovirus tn79 gene encodes a functional sulfhydryl oxidase enzyme that is able to support the replication of Autographa californica multiple nucleopolyhedrovirus lacking the sulfhydryl oxidase ac92 gene. Virology 460-461:207-216.